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題 名 | Primed in Situ (PRINS) Labeling for Rapid Detection of Numeric and Structural Chromosome Anomalies=利用引子原位標誌法快速診斷數目或構造上之染色體異常 |
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作 者 | 侯家瑋; 王作仁; | 書刊名 | 臺灣醫學會雜誌 |
卷 期 | 96:1 1997.01[民86.01] |
頁 次 | 頁46-50 |
分類號 | 417.133 |
關鍵詞 | 引子原位標誌法; 染色體異常; primed in situ (PRINS) labeling; Telomere; Ring chromosome; Supernumerary marker chromosome; |
語 文 | 英文(English) |
英文摘要 | Primed in situ (PRINS) labeling has been applied to replace the traditional fluorescence in situ hybridization (FISH) method for the detection of specific sequences in situ in several numerical and structural chromosome anomalies. PRINS is based on sequence-specific annealing in situ of an unlabeled DNA probe or oligonucleotide primer. The probe serves as a primer for chain elongation in situ, using the labeled nucleotides as substrate. An oligonucleotide, (CCCTAA) �f, representing human telomeric sequences, was mixed with nucleotides, biotin-16-dUTP, and Taq DNA polymerase, and applied on metaphase slides with ring chromosomes 4, 13, 18, X and Y. Primers for α -satellite sequences specific for the centromeric regions of human chromosomes 13, 15, 18, X and Y were also used to characterize the nature and origin of unidentifiable supernumerary marker chromosomes. The specificity of PRINS in differentiating centromeric sequences of chromosome 13 from 21, which is not possible with FISH, was demonstrated. Absence of the telomeric sequences in all of the ring chromosomes was noted in normal and abnormal phenotypes. The results suggest a mechanism of ring formation, an end-to-end fusion after loss of the palindromic nucleotide sequences at the telomeres. PRINS, a fast and sensitive method of detecting nucleic acid sequences in situ, may be a reliable technique for detecting chromosomal aneuploidies and some structural rearrangements. |
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